Long-term primary culture of epithelial cells from rainbow trout Oncorhynchus mykiss) liver.

Published

Journal Article

Long-term primary cultures of epithelial cells from rainbow trout (Oncorhynchus mykiss) liver have been established. Nearly homogenous (> 97%) populations of hepatocytes were placed into primary culture and remained viable and proliferative for at least 70 d. In addition to hepatocytes, proliferative biliary cells persisted in the cultures for at least 30 d. Finally, a third type of epithelial cell, which we have termed a "spindle cell," consistently appeared and proliferated to confluence in these cultures. The confluent cultures of spindle cells were successfully subcultured and passaged. The initial behavior, growth, and optimization of serum and media requirements for these cells is described. All three cell types proliferated as measured by thymidine incorporation, autoradiography, proliferating cellular nuclear antigen analysis, and propidium iodine staining. Further efforts to characterize the cells included western blotting and immunohistochemical staining with antibodies to cytokeratins previously reported in fish liver. From these data, it appears that all three cell populations are epithelial in nature. Furthermore, significant changes in actin organization, often indicative of transformation or pluripotent cells, were observed with increased time in primary culture.

Full Text

Duke Authors

Cited Authors

  • Ostrander, GK; Blair, JB; Stark, BA; Marley, GM; Bales, WD; Veltri, RW; Hinton, DE; Okihiro, M; Ortego, LS; Hawkins, WE

Published Date

  • May 1995

Published In

Volume / Issue

  • 31 / 5

Start / End Page

  • 367 - 378

PubMed ID

  • 7543343

Pubmed Central ID

  • 7543343

Electronic International Standard Serial Number (EISSN)

  • 1543-706X

International Standard Serial Number (ISSN)

  • 1071-2690

Digital Object Identifier (DOI)

  • 10.1007/bf02634286

Language

  • eng